2011年度神奈川大学総合理学研究所共同研究助成論文新規蛍光性物質AcO-TPKEを用いたウニ胚の染色について

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原著　2011 年度神奈川大学総合理学研究所共同研究助成論文New fluorescent substrates benzothiazolylphenol–substituted ketoester（TPKE）and acetoxyl‐TPKE（AcO‐TPEK）are possible indicators for living organisms. Both substrates stained the mid‐piece that was a mitochondrion in sea urchin sperm． AcOTPKE showed a higher rate of relative intensity than TPKE．The fluorescent intensity of AcO‐TPKE was dependent on enzymatic hydrolysis，catalyzed by esterase． Enzymatic analyses of the hydrolysis of AcO‐TPKE suggested that activity in extracts settled from the unfertilized stage to cleavage stages in sea urchins, but changed after the developmental stage of blastula or hatching．After staining fertilized eggs with AcO‐TPEK，they were allowed to develop normally and it was observed that the whole eggs and embryos emitted fluorescent signals；however，part of the body of larvae showed signals after the gastrula stage．In particular，pluteus larvae showed a strong signal in digestive organs．This may be related to the differentiation of digestive organs or the expression of digestive enzymes．According to these results，AcO‐TPKE is a possible fluorescent probe for living organisms，and esterase activities change throughout sea urchin development．